Distribution And Expression Of Peroxiredoxin Ⅰ And Ⅱ In Oviduct And Uterus And Effects Of Peroxiredoxin Ⅱ On The Development Of Mouse Preimplantation Embryo

Objective:To investigate the distribution of Peroxiredoxinâ… andâ…¡(Prxâ… andâ…¡)and the changes of their expression levels in the mouse oviduct and uterus during the estrous cycle. And to investigate the effects of Prxâ…¡on the development of mouse preimplantation embryo.Methods:1. Immunohistochemistry was used to observe the distribution of Prxâ… andâ…¡in the mouse oviduct and uterus.2. Western blotting was applied to detected the changes of Prxâ… andâ…¡expression levels in the mouse oviduct and uterus.3. KM 1-cell embryos were cultured in M16 media which were added different concentrations of Prxâ…¡protein. The effects of Prxâ…¡on the development of preimplantation embryo in vitro were observed, and the percentage of embryos developing from 2- to 4-cell embryos was used as evaluation criteria.4. KM and B6C3F1 1-cell embryos were cultured in M16 media. The different development capacity was observed. The percentages of embryos developing from 2- to 4-cell embryos and to blastocysts were used as evaluation criteria.5. With redox-sensitive fluorescence probe 2', 7'-dichlorofluorescein diacetate (DCFH-DA), the differences of reactive oxygen species (ROS) were monitored between KM 2-cell embryos developed in vitro and in vivo by laser confocal scanning microscopy, as well as the differences of ROS level between KM and B6C3F1 2-cell embryos developed in vitro.Results: 1. In the mouse oviduct, immunopositivity for both Prxâ… andâ…¡was found in oviductal epithelia; in the mouse uterus, immunopositivity for Prxâ… was observed in uterine endometrial epithelia, glandular epithelia and endometrial stroma, while Prxâ…¡was seen mainly in endometrial stroma.2. Western blotting showed that Prxâ… andâ…¡were expressed in the uterus and oviduct, and their levels of expression had no significant differences between diestrus and estrus.3. KM 1-cell embryos were cultured in M16 media which were added by different concentrations of Prxâ…¡protein. The percentages of embryos developing from 2- to 4-cell embryos had no significant differences comparing to the control.4. Most of KM 1-cell embryos were arrested in 2-cell stages when cultured in M16 media, only seldom of them could develop to blastocysts. While most of B6C3F1 1-cell embryos could develop over 2-cell stage and to blastocyst in vitro.5. ROS level of KM 2-cell embryos developed in vitro was significant less than those developed in vitro. And it also was less than B6C3F1 2-cell embryos developed in vitro, but there was no significant difference between them.Conclusion:Prxâ… andâ…¡expressed stably in the mouse oviduct and uterus during the estrous cycle, and they might supply a good antioxidant microenvironment for fertilization and embryo development. But the M16 media with Prxâ…¡could not improve the development of KM mouse preimplantation embryo in vitro, and could not overcome the 2-cell block either. The reason might be that ROS level of KM 2-cell embryos developed in vitro was not increased. So 2-cell block of KM mouse embryos in vitro was not mainly caused by the increase of ROS level.